Eucommia ulmoides extract ameliorates sarcopenia by activating the GH/IGF-1 signaling axis

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Raw data for EUE ameliorates sarcopenia

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The research data were gathered through an integrated pipeline of in silico discovery and multi-dimensional in vivo validation. 1. Bioinformatics & Network Pharmacology: Transcriptomic datasets (GSE285869, GSE227495, GSE269698) were obtained from the GEO repository and analyzed using R software (v4.3.1) with DESeq2 and limma packages. Functional enrichment (GO/KEGG) was performed using the clusterProfiler package. Network pharmacology (TCMSP, SwissTargetPrediction, GeneCards, OMIM) and STRING (confidence ≥0.7) identified core targets such as AKT1, MTOR, and IGF1R, which were further validated via molecular docking. 2. EUE Preparation & HPLC: Eucommia bark extract (EUE) was prepared using enzymatic hydrolysis (0.1% cellulase, 50°C, pH 5.0) followed by ultrasonic-assisted extraction (40 kHz, 500 W). Chemical constituents (aucubin, geniposidic acid, chlorogenic acid, and geniposide) were identified and quantified using an Agilent 1260 HPLC system with a ZORBAX Eclipse Plus C18 column. 3. Animal Modeling: Sarcopenia was induced in male BALB/c mice through combined treatment with D-galactose (100 mg/kg/day) and hydrocortisone (35 mg/kg/day). Mice were treated with EUE (500–2000 mg/kg) or the selective IGF-1R inhibitor Picropodophyllin (PPP). 4. Functional Assessment: Motor capacity was measured via grip strength (normalized to body weight), weighted swimming exhaustion, and rotating rod fatigue tests (uniform acceleration from 4 to 40 r/min). 5. Biochemical Assays: Serum and tissue levels of AGEs, GH, and IGF-1 were quantified using ELISA kits. MDA levels were measured via the TBA method, with absorbance recorded on a BioTek microplate spectrophotometer. 6. Histology & Ultrastructure: Muscle sections were stained with H&E for cross-sectional area (CSA) or IHC for fiber-typing (Fast/Slow Myosin Heavy Chain antibodies). Organelle integrity was assessed via TEM (JEM-1400 FLASH) and evaluated using the Flameng Score. All quantification was performed using ImageJ software. 7. Molecular Biology: Western Blotting analyzed the phosphorylation of the STAT5b-IGF-1R-Akt-mTOR-FoxO3a cascade using RIPA buffer, SDS-PAGE, and PVDF membranes, with densitometry performed via Image Studio Lite. qRT-PCR quantified mRNA levels (e.g., Myog, Myod, Mstn, Redd1, Pgc1a, Bnip3) using TRI pure Reagent, SYBR Green, and the 2⁻ΔΔCt method. 8. Statistical Analysis: Data were analyzed in R (v4.4.1) using one-way ANOVA followed by Tukey’s post-hoc test for continuous data and the Wilcoxon rank-sum test for pathological scores.

Institutions

• Xihua University

  Sichuan, Chengdu

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